Synthesis, physicochemical characterization, and crystallization of a putative retro-coiled coil

OBJECTIVES: The purposes of this study were to investigate the use of radial artery applanation tonometry and a generalized transfer function for the assessment of central aortic pressure augmentation in subjects taking commonly used antihypertensive agents (angiotensin-converting enzyme inhibitors, beta-adrenergic blockers, Ca2+ antagonists, diuretic therapy). BACKGROUND: Applanation tonometry of the radial artery with a generalized transfer function has been proposed as a means of assessing central aortic blood pressure. Recently, a commercial apparatus based on this technique has become available; we therefore examined the effect of a generalized transfer function on derived central aortic pressure compared with measured brachial blood pressures and also investigated the potential of this technique to assess the influence of differing drug therapy. METHODS: Two hundred and sixty-two hypertensive patients on stable medication were studied using the PWV Medical Blood Pressure Analysis System (version 2, DAT-1). RESULTS: In univariate analysis, augmentation index showed association with age, sex, height and heart rate. In multivariate analysis, diastolic blood pressure and age (positively), height and heart rate (negatively) and sex were significantly associated. After adjustment for these variables, pressure augmentation was not associated with any antihypertensive treatment investigated. Linear relationships were demonstrated between brachial blood pressures and corresponding central pressures derived by transfer function methods. CONCLUSIONS: Our findings suggest that if adjustment for central-peripheral pressure difference is necessary, simple linear relationships may be sufficient. Age, heart rate and height but not the class of antihypertensive medication affected the degree of pressure augmentation observed using this technique.     

Fracture healing stimulator based on pulsed electromagnetic fields and its clinical application

Two large epidermoid cysts of the spleen have been accidentally discovered in two young men. These cysts are uncommon and represent about 10% of the non parasitic benign cysts of the spleen. From these cases, the authors reviewed the literature. These cysts, which are caused by an abnormal development during the seventh week of the intra-uterine life, are often asymptomatic but the occurrence of a complication may enable the diagnosis. This diagnosis relies on the surgical ablation of the cyst and the histopathologic examination. The surgery should be as conservative as possible.     

Recombinant human luteinizing hormone: a partial physicochemical, biological and immunological characterization

The aim of this study was to partially characterize the glycoform composition of a recombinant human luteinizing hormone preparation (rhLH; Serono), an early version of the material (LHadi) which is currently being assessed for clinical application. Specifically, the charge (pl) and internal carbohydrate complexity of this rhLH was examined and compared with that of an alternative commercially available form of recombinant LH (Crystal Chem) and a pituitary International Reference Preparation (IRP). All preparations were separated by charge by chromatofocusing them on a pH gradient (7-4) using a 4 ml mono-P column is conjunction with a fast performance liquid chromatography system and by complexity of the oligosaccharide structures using concanavalin A (con-A) lectin affinity chromatography. LH in both the unfractionated and fractionated material was assessed by immunoradiometric assay (IRMA, I-LH) and by the in-vitro Leydig cell bioassay (B-LH). Both assays were calibrated against IRP 80/552. The in-vitro biopotency of the preparations was 18187 (Serono rhLH), 12063 (Crystal Chem rhLH) and 6658 (80/552) IU/mg; biological:immunological ratios were 1.14 (80/552), 1.90 (Crystal Chem rhLH) and 1.99 (Serono rhLH). However, similar qualitative data were obtained by both bioassay and immunoradiometric assay following fractionation, with the median pl of the bioactive LH in the preparations being 5.5 (24% > pH 6), 5.52 (18% > pH 6) and 4.97 (0% > pH 6) for the Serono, Crystal Chem and pituitary preparations respectively. Further all three contain < 1% of the complex carbohydrate structures and between 36-44% and 56-63% of the intermediate and simple forms of bioactive LH. In conclusion, the Serono recombinant LH preparation has a higher in-vitro bioactivity and is more basic than the other two preparations although the complexity of its carbohydrate moities appears to be similar.     

Mechanistic studies of the biosynthesis of paratose: purification and characterization of CDP-paratose synthase

The 3,6-dideoxyhexoses can be found in the cell wall lipopolysaccharide of Gram-negative bacteria, where they have been shown to be the dominant antigenic determinants. All naturally occurring 3,6-dideoxyhexoses, with colitose as the only exception, are biosynthesized via a complex pathway that begins with CDP-d-glucose. Included in this pathway is CDP-paratose synthase, an essential enzyme in the formation of the 3,6-dideoxy sugars, CDP-paratose and CDP-tyvelose. Recently, the gene encoding CDP-paratose synthase in Salmonella typhi, rfbS, has been identified and sequenced [Verma, N., and Reeves, P. (1989) J. Bacteriol. 171, 5694-5701]. On the basis of this information, we have amplified the rfbS gene by polymerase chain reaction (PCR) from S. typhi and cloned this gene into a pET-24(+) vector. Expression and purification of CDP-paratose synthase have allowed us to fully characterize the catalytic properties of this enzyme, which is a homodimeric protein with a preference for NADPH over NADH. It catalyzes the stereospecific hydride transfer of the pro-S hydrogen from the C-4' position of the reduced coenzyme to C-4 of the substrate, CDP-3,6-dideoxy-D-glycero-D-glycero-4-hexulose. The overall equilibrium of this catalysis greatly favors the formation of the reduced sugar product and the oxidized coenzyme. Interestingly, this enzyme also exhibits a high affinity for NADPH with a much smaller dissociation constant (Kia) of 0.005 +/- 0.002 microM compared to the Km of 26 +/- 8 microM for NADPH. While this unusual property complicated the interpretation of the kinetic data, the kinetic mechanism of CDP-paratose synthase as explored by the combination of bisubstrate kinetic analysis, product inhibition studies, and dead-end competitive inhibition studies is most consistent with a Theorell-Chance mechanism. The present study on CDP-paratose synthase, a likely new member of the short-chain dehydrogenase family, represents the first detailed characterization of this type of ketohexose reductase, many of which may share similar properties with CDP-paratose synthase.     

Microprocessor based spatial TENS (transcutaneous electric nerve stimulator) designed with waveform optimality for clinical evaluation in a pain study

1. The use of organs from animal donors (xenotransplantation) is a potential solution to the chronic shortage of allogeneic organs and currently the pig is thought to be the most suitable donor for man. However, porcine organs are rejected rapidly by a vascular process called hyperacute rejection which has so far prevented clinical xenotransplantation. Although it is likely that this barrier will be overcome in the near future by the application of novel strategies, probably involving the use of organs from transgenic pigs, data from animal models indicate that multiple other immune mechanisms will contribute to the rejection of xenografts. 2. We have described two aspects of these immune mechanisms. First, the phenomenon of 'accommodation', whereby xenografts acquire in vivo resistance to vascular rejection, has been explored in an in vitro model utilizing immortalized porcine endothelial cells. The results indicate that human anti-pig antibodies induce a concentration-dependent and time-dependent change in porcine endothelial cells compatible with the development of accommodation. 3. Secondly, the in vitro human anti-porcine T-cell response has been documented in detail, with particular emphasis on quantitative and qualitative comparisons with the in vitro T-cell alloresponse. The results of this work, which indicate that the response to porcine xenografts is likely to be significantly stronger than that against allografts, have important implications for the level of conventional immunosuppression that may be necessary to prevent xenograft rejection, and provide an important basis for the development of strategies to promote xenograft-specific immunosuppression and tolerance.     

Isolation and structural identification of bifidogenic growth stimulator produced by Propionibacterium freudenreichii

A bifidogenic growth stimulator produced by Propionibacterium freudenreichii was purified, and its chemical structure was determined. We obtained 7.1 mg of a bifidogenic growth stimulator from 1738 g of lyophilized P. freudenreichii cells by silica gel column chromatography, Sephadex LH-20 column chromatography, and preparative HPLC. The mass of the bifidogenic growth stimulator was 217.037 (C11H7NO4) as determined by high resolution mass spectrometry. Various experimental analyses indicated that the chemical structure of the bifidogenic growth stimulator was 2-amino-3-carboxy-1,4-naphthoquinone.     

基于冶金物理化学原理推动冶金企业智能制造发展

依据公开文献报道讨论目前宝钢股份智能化生产发展概况,基于生产数据及冶金理论,针对宝钢产品中夹杂物的控制,从转炉、精炼、连铸等重点工序展开,讨论了转炉造渣控制、精炼OB(Oxygen blowing)控制,以及连铸的吹氩控制.基于冶金理论重点阐述了精炼吹氧脱碳、连铸中间包吹氩对液态钢水中氧的影响,阐明钢铁工业中工艺数据与冶金原理密切结合起来,才能推动冶金工业智能制造更好的发展.     

Mechanistic study on liver tumor promoting effects of piperonyl butoxide in rats

Piperonyl butoxide, alpha-[2-(2-butoxyethoxy)ethoxy]-4,5-methylenedioxy-2-propyltol uene, is a widely used pesticide-synergist. Recently, results were reported indicating that piperonyl butoxide is a hepatocarcinogen in rat. Since the underlying mechanism was not elucidated, we examined the effects on rat liver cells in detail. For this purpose male F344 rats were administered piperonyl butoxide mixed in the diet at concentrations of 0 (negative control), 0.05, 0.2 or 2% for 2 days, 1, 2, and 4 weeks. As a positive control, phenobarbital was administered to rats for up to 4 weeks as a 0.1% solution in the drinking water. Increased liver weight, centrilobular hepatocellular hypertrophy due to increased smooth endoplasmic reticulum, decreased numbers and areas of connexin 32-positive spots per hepatocyte, and increased cell proliferation were observed in rats treated with 0.2 and 2% piperonyl butoxide. Similar results were obtained for 0.1% phenobarbital treated rats. Hepatocellular necrosis suggestive of hepatotoxicity was also observed in the 2% piperonyl butoxide group. These results indicate that the promoting mechanism of piperonyl butoxide in hepatocarcinogenesis is similar to that of phenobarbital, involving an ability to induce CYP isoenzymes and inhibit gap junctional intercellular communication. In addition, increased cell proliferation following hepatocellular necrosis may also play a role at high doses.     

Effect of serum proteins and osteoblasts on the surface transformation of a calcium phosphate coating: a physicochemical and ultrastructural study

We describe a practical method for the analysis of multiple analytes in a single sample. The vehicle for each separate measurement consists of a set of microspheres identifiable by characteristic fluorophores embedded in the particles. The use of robust, bench-top flow cytometers (flow microfluorimeters) for the analysis of the multiple sets of microspheres is facilitated by hardware and software, which acquire the data from the cytometer, classify the microspheres according to sets, and collate measurement information from each microsphere set in real time. This measurement system can analyze up to 64 analytes in a single sample. The advantages of multiplexed assays using flow cytometry include robust measurements, because each microsphere set is measured repeatedly. The advantage of the assay's is consistent with simultaneous measurement of many parameters as well as the speed with which the flow microfluorimeter (cytometer) makes measurements (many hundreds per second). Here, we describe the properties of the microspheres, the calibration of the cytometer, and the influence of the properties of the microspheres on the sensitivity of measurements.     

Purification, physicochemical characterization and biological properties of a lectin from Erythrina velutina forma aurantiaca seeds

A lectin was purified from seeds of Erythrina velutina forma aurantiaca by affinity chromatography on cross-linked guar gum. The lectin is a potent agglutinin for human (minimal concentration of protein able to cause visible agglutination of a 2% erythrocyte suspension varying from 1 to 4 micrograms/ml), rabbit (4 micrograms/ml) and chicken erythrocytes (8 micrograms/ml) but presented low activity against cow (250 micrograms/ml) or sheep (333 micrograms/ml) blood cells. Hemagglutination of human O+ erythrocytes was inhibited by D-lactose (0.2 mM) > D-galactose (0.8 mM) > D-raffinose (2.1 mM). At pH 7.5, chromatography on a Superose 12 HR 10/30 column showed that the lectin was primarily a dimer (56.0 kDa) composed of two identical subunits (31.6 kDa each). A small amount of a tetrameric form was also apparently present. The lectin is a glycoprotein (7.3% carbohydrate), has a pI of 4.5, contains high levels of acidic (Asp and Glu, 64.2 and 51.6 residues/mol, respectively) and hydroxy amino acids (Ser and Thr, 42.9 and 38.5 residues/mol, respectively) but relatively low amounts of sulfur amino acids (Cys and Met, 1.0 and 5.0 residues/mol, respectively) and has an N-terminal sequence of Val-Glu-Thr-Ile/Leu-Pro-Phe-Ser. Its hemagglutinating activity was abolished by heating at 70 degrees C for 10 min. The activation energy (delta G') required for denaturation measured by loss of hemagglutination activity was 24.87 kcal/mol. In rats, the purified lectin (100 micrograms) induced neutrophil migration into the peritoneal cavity (3.7 +/- 0.6 x 10(6) neutrophils/ml) or into the air pouch (2.75 +/- 0.25 x 10(6) neutrophils/ml), 8 and 10 times greater than the negative control, respectively.     

Development of ciprofloxacin-loaded nanoparticles: physicochemical study of the drug carrier

This paper describes the optimization of the preparation of ciprofloxacin-loaded polyethylbutylcyanoacrylate (PEBCA) nanoparticles. The association of ciprofloxacin with nanoparticles was performed by emulsion polymerization, but successful entrapment was only obtained in the presence of acetone in the polymerization medium. This preparation process led to a stable ciprofloxacin nanoparticle suspension, with a mean size value twice as high as that obtained in the absence of drug, and an association efficiency of 82%. Moreover, the molecular weight value of ciprofloxacin nanoparticles was shown to be reduced as compared with unloaded nanoparticles. Drug release from the colloidal carrier in medium containing esterase was found to be very slow (a maximum of 51.5% after 48 h), suggesting that this release resulted from bioerosion of the polymer matrix. Interestingly, it was observed that 30.5% of the initial amount of ciprofloxacin was not detectable by HPLC analysis after nanoparticle preparation and corresponded either to ciprofloxacin covalently bound to PEBCA or to ciprofloxacin chemically degraded during the polymerization process. 19F-NMR analysis demonstrated that ciprofloxacin entrapped into nanoparticles was only in its neutral form. The measurements of molecular weight suggest the participation of the antibiotic as an anionic polymerization initiator, leading to the formation of a chemical bond between some of the drug and the polymer. These data allowed us to propose a model describing the association of ciprofloxacin with PEBCA nanoparticles obtained by emulsion polymerization.     

A comparison of phorbol myristate acetate with phytohemgglutinin as a stimulator of in vitro colony growth of peripheral human leucocytes

Peripheral blood mononuclear cells from normal individuals were cultured in soft agar (0.3%) with either phytohaemagglutinin (PHA) or the tumor promoter phorbol myristate acetate (PMA). PMA was found to induce colony growth under these conditions as well as that achieved by PHA. The colonies from PHA stimulation were entirely lymphoid but with PMA the type of cell was more varied with some cells that appeared to be of the granulocytic-macrophage line. PMA thus appears to have the ability to promote growth in cells other than lymphocytes in the peripheral blood under these conditions.     

Studies on the production and action of polymorph migration stimulator

Human blood monocytes are known to react with hydrocortisone in vitro to produce a factor which stimulates polymorph migration. This study shows that the polymorph migration stimulator (PMS) is generated only by steroids with glucocorticoid activity. The interaction between steroid and monocytes is dependent on protein synthesis. Serum is necessary for the expression of the activity of the factor, but the steroid-monocyte reaction can occur in the absence of serum. The stimulatory effect on migration appears to be specific for polymorphs and does not affect mono-nuclear leucocytes. In addition to blood monocytes, spleen and bone marrow cells also react with corticosteroids to produce the polymorph migration stimulator.     

Identification and characterization of a novel member of the fibroblast growth factor family

A new member of the fibroblast growth factor (FGF) family, FGF-13, has been molecularly cloned as a result of high throughput sequencing of a human ovarian cancer cell library. The open reading frame of the novel human gene (1419 bp) encodes for a protein of 216 a.a. with a molecular weight of 22 kDa. The FGF-13 sequence contains an amino-terminal hydrophobic region of 23 a.a. characteristic of a signal secretion sequence. FGF-13 is most homologous, 70% similarity at the amino acid level, to FGF-8. Northern hybridization analysis demonstrated prominent expression of FGF-13 in human foetal and adult brain, particularly in the cerebellum and cortex. In proliferation studies with BaF3 cells, FGF-13 preferentially activates cell clones expressing either FGF receptor variant, 3-IIIc or 4. The signal transduction pathways of FGF-13 and FGF-2 were compared in rat hippocampal astrocytes. The two FGFs induce an equivalent level of tyrosine phosphorylation of mitogen-activated protein kinase (MAPK) and c-raf activation. However, FGF-13 is more effective than FGF-2 in inducing the phosphorylation of phospholipase C-gamma (PLC-gamma). Treatment of neuronal cultures from rat embryonic cortex with FGF-13 increases the number of glutamic acid decarboxylase immunopositive neurons, the level of high-affinity gamma-aminobutyric acid (GABA) uptake, and choline acetyltransferase enzyme activity. The GABAergic neuronal response to FGF-13 treatment is rapid with a significant increase occurring within 72 h. We have identified a novel member of the FGF family that is expressed in the central nervous system (CNS) and increases the number as well as the level of phenotypic differentiation of cortical neurons in vitro.     

Bovine placental lactogen is a potent stimulator of growth and displays strong binding to hepatic receptor sites of coho salmon

Juvenile coho salmon were treated with bovine placental lactogen (bPL) and bovine growth hormone (bGH) to examine the growth promoting activities of these proteins in a lower vertebrate. Fish were intraperitoneally injected either with 0.5 or 5.0 micrograms/g bPL or with 5.0 micrograms/g bGH once a week for 5 weeks. After only a single injection and 1 week of growth, the high dose of bPL stimulated a significant increase in weight and length relative to untreated fish or fish treated with a control protein, bovine serum albumin. At the end of the experiment, all hormone-treated groups were significantly larger than controls. Fish treated with 5 micrograms/g bPL gained more than three times as much weight as controls. The 5.0 micrograms/g bGH group grew at the same rate as fish treated with one-tenth this dose of bPL, indicating that bPL is a potent stimulator of growth in this species. Radioreceptor assays performed on coho salmon liver membrane preparations indicate that bPL binds with approximately 430-fold higher affinity than bGH, and some 8000-fold higher affinity than bovine prolactin. The action of bPL relative to the structure and function of salmonid pituitary hormones is discussed.     

基于COD组分表征的GPS-X对污水厂的模拟

介绍了活性污泥系统仿真软件GPS-X的模型机理及其建模的过程,结合COD组分表征结果,通过实例分析了该软件在污水处理厂中模拟应用,模拟结果为:COD最大误差为5.48%,最小误差为0.83%,相对误差绝对平均值约2.85%;TN最大误差为7.23%,最小误差为0.15%,相对误差绝对平均值约3.62%,NH+4-N最大误差为9.17%,最小误差为1.05%,相对误差绝对平均值约4.98%,模拟结果与实测结果趋势吻合一致,能较好地反映污水处理厂实际状况.     

Characterization of a guanine nucleotide dissociation stimulator for a ras-related GTPase

ras-related GTPases participate in signaling for a variety of cellular processes. The GTPases cycle between a GTP-bound active state and a GDP-bound inactive state. This cycling is partially controlled by guanine nucleotide dissociation stimulators (GDS, also known as exchange factors). We report on the molecular cloning of cDNAs encoding a new mammalian GDS protein, using sequences derived from the yeast ras GDS proteins as probes. The encoded protein stimulates the dissociation of guanine nucleotides from the ras-related ralA and ralB GTPases at a rate at least 30-fold faster than the intrinsic nucleotide dissociation rate. This new GDS, ralGDS, is at least 20-fold more active on the ralA and ralB GTPases than on any other GTPase tested, including other members of the ras family (H-ras, N-ras, K-ras, R-ras, rap1a and rap2), members of the rho family (rhoA, rhoB and CDC42-Hs) and members of the rab family (rab3a and ypt1). While the ralGDS protein is phosphorylated on serine residues, we find no evidence that phosphorylation affects the activity of insect cell-expressed ralGDS towards the ralA or ralB GTPase. The 3600 nucleotide ralGDS mRNA and the 115 kDa protein were found in all tissues and cell lines examined.     

Optimizing the technology for making automobile sheet 08Yu based on the physicochemical principles of ladle metallurgy

The main method of eliminating the possibility of having the steel contaminated by nonmetallic inclusions is optimizing the parameters of the ladle treatment on the basis of reliable physicochemical representations of the processes of refining, alloy, and finishing of the steel, the interactions of the metal and slag phases, and other aspects of ladle metallurgy. This article examines the main steps being taken in this direction at the Cherepovets Metallurgical Combine Severstal’ in the production of high-quality automobile sheet 08Yu. __________ Translated from Metallurg, No. 8, pp. 58–65, August, 2007.